Journal of Food Hygiene and Safety (한국식품위생안전성학회지)

The Korean Society of Food Hygiene and Safety (한국식품위생안전성학회)
권호 표지
DOI QR코드

DOI QR Code

Analytical Method for Determination of Cephalexin in Bovine Edible Tissues using Liquid Chromatography Coupled to Tandem Mass Spectrometry

LC-MS/MS를 이용한 소의 식용조직 중 세팔렉신의 잔류검사법

  • Chae, Won-Seok (Department of Chemistry, Daejin University) ;
  • Lee, Sung Joong (Department of Chemistry and Research Institute of Live Science, Gyeongsang National University) ;
  • Son, Song-Ee (Institute of Animal Medicine and College of Veterinary Medicine, Gyeongsang National University) ;
  • Kim, Suk (Institute of Animal Medicine and College of Veterinary Medicine, Gyeongsang National University) ;
  • Lee, Hu-Jang (Institute of Animal Medicine and College of Veterinary Medicine, Gyeongsang National University)
  • 채원석 (대진대학교 자연과학대학 화학과) ;
  • 이성중 (경상대학교 화학과.생명과학연구원) ;
  • 손송이 (경상대학교 수의과대학.동물의학연구소) ;
  • 김석 (경상대학교 수의과대학.동물의학연구소) ;
  • 이후장 (경상대학교 수의과대학.동물의학연구소)
  • Received : 2017.11.11
  • Accepted : 2017.12.16
  • Published : 2018.02.28
Download PDF
⟨ Previous Next ⟩

Abstract

An analytical method for the determination of cephalexin (CEX) in bovine tissues (muscle, liver, kidney and fat tissues) was developed and validated using liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS). Tissue samples were extracted by the liquid-liquid extraction based on 5% trichloroacetic acid (TCA). The chromatographic separation was achieved on a reverse phase $C_{18}$ column with gradient elution using a mobile phase of 20 mM hexafluroacetylacetone (HFAC)/50% acetonitrile (40:60). The procedure was validated according to the Ministry of Food and Drug Safety guideline determining accuracy, precision, and limit of detection. Mean recoveries of CEX from spiked edible tissues ($6{\sim}1,500{\mu}g/kg$) were 83.9~106.8%, and the relative standard deviation was between 2.3 and 14.8%. Linearities were obtained with the correlation coefficient ($r^2$) of > 0.999. Limit of detection and limit of quantification for the investigated CEX were 2~10 and $6{\sim}30{\mu}g/kg$, respectively. This method was reliable, sensitive, economical and suitable for routine monitoring of CEX residues in bovine edible tissues.

본 연구는 소의 가식부위(근육, 신장, 간장, 지방) 중에서 세팔렉신을 효과적으로 정량분석하기 위한 LC-MS/MS법을 확립하고 이를 검증하기 위해 수행되었다. 확립된 LC-MS/MS에 대해 특이성, 검출한계, 정량한계, 정확도 및 정밀도에 대한 검증을 통하여 유효성을 확인하였다. 표준 용액을 이용하여 검량성을 작성한 결과, $r^2$ > 0.999 이상의 직선성을 나타내었으며, 세팔렉신에 대한 검출한계와 정량한계는 각각 2~10과 $6{\sim}30{\mu}g/kg$으로 나타났다. 또한, 회수율은 83.9~106.8%로 나타났으며, 상대표준편차는 2.3~14.8%로 나타나 정확성이 우수하였다. 이는 식품의약품안전처의 잔류동물용의약품 분석법에서 제시한 기준에 모두 적합한 수준이었다. 따라서 본 연구를 통해 개발된 LC-MS/MS법은 향후 소의 가식부위 중 세팔렉신을 분석하는데 효과적으로 활용될 수 있을 것으로 사료된다.

Keywords

References

  1. Mainero Rocca, L., Gentili, A., Perez-Fernandez, V. and Tomai, P.: Veterinary drugs residues: a review of the latest analytical research on sample preparation and LC-MS based methods. Food Addit. Contam. Part A Chem. Anal. Control Expo. Risk Assess. 34, 766-784 (2017).
  2. Farouk, F., Azzazy, H.M. and Niessen, W.M.: Challenges in the determination of aminoglycoside antibiotics, a review. Anal. Chim. Acta. 890, 21-43 (2015). https://doi.org/10.1016/j.aca.2015.06.038
  3. Moreno-Bondi, M.C., Marazuela, M.D., Herranz, S. and Rodriguez, E.: An overview of sample preparation procedures for LC-MS multiclass antibiotic determination in environmental and food samples. Anal. Bioanal. Chem. 395, 921-943 (2009).
  4. Sawant, A.A., Sordillo, L.M. and Jayarao, B.M.: A survey on antibiotic usage in dairy herds in pennsylvania. J. Dariy Sci. 88, 2991-2999 (2005). https://doi.org/10.3168/jds.S0022-0302(05)72979-9
  5. Fisher, J.F., Meroueh, S.O. and Mobashery, S.: Bacterial resistance to beta-lactam antibiotics: compelling opportunism, compelling opportunity. Chem. Rev. 105, 395-424 (2005). https://doi.org/10.1021/cr030102i
  6. Drawz, S.M. and Bonomo, R.A.: Three decades of beta-lactamase inhibitors. Clin. Microbiol. Rev. 23, 160-201 (2010). https://doi.org/10.1128/CMR.00037-09
  7. Ministry of Food and Drug Safety (MFDS).: Korean Food Standards Codex. Appendix 6: Maximum residue limits of veterinary drugs in food. MFDS, Cheongju, Korea, pp. 21 (2017).
  8. The European Agency for the Evaluation of Medical Products.: Committee for veterinary medicinal products cephalexin summary report: EMEA/MRL/627/99-FINAL. Veterinary Medicines Evaluation Unit, London, U.K., pp. 8 (1999).
  9. Panda, S.S., Ravi, K., Bera, V.V., Dash, R. and Mohanta, G.: Determination of cephalexin monohydrate in pharmaceutical dosage form by stability-indicating RP-UFLC and UV spectroscopic methods. Sci. Pharm. 81, 1029-1036 (2013). https://doi.org/10.3797/scipharm.1306-07
  10. Jeswani, R.M., Sinha, P.K., Topagi, K.S., Damle, M.C.: A validated stability indicating HPTLC method for determination of cephalexin in bulk and pharmaceutical formulation. Int. J. Pharm. Tech. Res. 1, 527-536 (2009).
  11. Edward, P.C. and Stanley, G.W.: Molecularly imprinted solid phase extraction for rapid screening of cephalexin in human plasma and serum. Anal. Chim. Acta. 481, 165-174 (2003).
  12. Argekara, A.P., Raja, S.V. and Kapadiaa, S.U.: Simultaneous determination of cephalexin and carbocisteine from capsules by reverse phase high performance liquid chromatography (RP-HPLC). Anal. Lett. 30, 821-831 (1997). https://doi.org/10.1080/00032719708006427
  13. Regina, V.O., Angela, C.D. and Quezia, B.C.: Quantification of cephalexin as residue levels in bovine milk by high-performance liquid chromatography with on-line sample cleanup. Talanta. 71, 1233-1238 (2007). https://doi.org/10.1016/j.talanta.2006.06.024
  14. Kim, M.R., Choi, Y.H., Choi, H., Kim, D.H., Kim, Y.S. and Lee, J.H.: Monitoring for cephalosporins residues in raw meat in Seoul. Korean J. Vet. Serv. 38, 259-264 (2015).
  15. Li, W., Shen, H., Hong, Y., Zhang, Y., Yuan, F. and Zhang, F.: Simultaneous determination of 22 cephalosporins drug residues in pork muscle using liquid chromatography-tandem mass spectrometry. J. Chromatogr. B Analyt. Technol. Biomed. Life Sci. 1022, 298-307 (2016). https://doi.org/10.1016/j.jchromb.2016.04.026
  16. Liu, X., Yu, Y., Zhao, M., Zhang, H., Li, Y. and Duan, G.: Solid phase extraction using magnetic core mesoporous shell microspheres with C18-modified interior pore-walls for residue analysis of cephalosporins in milk by LC-MS/MS. Food Chem. 150, 206-212 (2014). https://doi.org/10.1016/j.foodchem.2013.10.145
  17. Perez-Burgos, R., Grzelak, E.M., Gokce, G., Saurina, J., Barbosa, J. and Barron, D.: Quechers methodologies as an alternative to solid phase extraction (SPE) for the determination and characterization of residues of cephalosporins in beef muscle using LC-MS/MS. J. Chromatogr. B Analyt. Technol. Biomed. Life Sci. 899, 57-65. (2012). https://doi.org/10.1016/j.jchromb.2012.05.002
  18. Chen, H., Li, X. and Zhu, S.: Occurrence and distribution of selected pharmaceuticals and personal care products in aquatic environments: a comparative study of regions in China with different urbanization levels. Environ. Sci. Pollut. Res. Int. 19, 2381-2389 (2012). https://doi.org/10.1007/s11356-012-0750-2
  19. Lee, Y.J. and Lee, H.S.: Simultaneous determination of cefoxitin, cefuroxime, cephalexin and cephaloridine in plasma using HPLC and a column-switching technique. Chromatographia. 30, 80-84 (1990). https://doi.org/10.1007/BF02270453
  20. Pecavar, A., Smidovnik, A., Milivojevic, D. and ProSek, M.: A reversed phase high-performance liquid chromatographic method for determination of cephalexin in human plasma. J. High Resol. Chrornatogr. 20, 674-678 (1997).
  21. Ministry of Food and Drug Safety (MFDS).: Korean Food Standards Codex. Chapter 7. General analytical method, 5. Monitoring of veterinary drug residues in foods. MFDS, Cheongju (2016).
  22. Li, W., Shen, H., Hong, Y., Zhang, Y., Yuan, F. and Zhang, F.: Simultaneous determination of 22 cephalosporins drug residues in pork muscle using liquid chromatography-tandem mass spectrometry. J. Chromatogr. B. 1022, 298-307 (2016). https://doi.org/10.1016/j.jchromb.2016.04.026
  23. National Institute of Food and Drug Safety Evaluation (NIFDS).: Handbook of residue analytical methods for veterinary drugs in food. NIFDS, Cheongju, Korea, pp. 3-4 (2014).
  24. Ministry of Agriculture, Food and Rural Affairs (MAFRA), Animal and Plant Quarantine Agency (APQA) and National Institute of Food and Drug Safety Evaluation (NIFDS).: National antimicrobial usage and resistance monitoring in 2015 - livestock and animal products. MAFRA, Sejong, Korea, pp. 9-10 (2016).
  25. Heo, J.H., Cho, M.H., Lee, K.C., Park, M.N., Cho, E.J., Choi, M.S., Kim, C.H., Kang, J.B., Kim, E. and Kim, J.S.: An epidemiological study on the calves with clinical diarrhea in southern Gyeongnam. Korean J. Vet. Serv. 31, 305-313 (2008).
  26. Son, B., Kim, J., An, C., Lee, S. and Kim, B., Chung, D.: Simultaneous analysis of ${\beta}$-lactam antibiotics and ${\beta}$-blockers by LC-MS/MS. Anal. Sci. Technol. 29, 179-185 (2016). https://doi.org/10.5806/AST.2016.29.4.179
  27. Carretero, V., Blasco, C. and Pico, Y.: Multi-class determination of antimicrobials in meat by pressurized liquid extraction and liquid chromatography-tandem mass spectrometry. J. Chromatogr. A. 1209, 162-173 (2008). https://doi.org/10.1016/j.chroma.2008.09.011
  28. The European Agency for the Evaluation of Medical Products (EMEA).: Cefalexin - summary report. Committee for veterinary medicinal products. Veterinary Medicines Evaluation Unit (EMEA/MRL/627/99) (1999).
  29. Codex guidelines for the establishment of a regulatory programme for control of veterinary drug residues in foods (CAC/GL 16) (1993).
  30. Validation guidelines, Codex committee on residues of veterinary drugs in foods, 11th session (1998).